U.S. patent number 4,714,612 [Application Number 06/748,393] was granted by the patent office on 1987-12-22 for composition for oral application.
This patent grant is currently assigned to Lion Corporation. Invention is credited to Tatsuo Kiyoshige, Tsuneaki Nakamura, Shuji Sasaki.
United States Patent |
4,714,612 |
Nakamura , et al. |
December 22, 1987 |
Composition for oral application
Abstract
A composition for oral application comprises a nonspecific
.gamma.-globulin as an effective ingredient for preventing
Bacteroides gingivalis from colonizing in the mouth.
Inventors: |
Nakamura; Tsuneaki (Odawara,
JP), Kiyoshige; Tatsuo (Hadano, JP),
Sasaki; Shuji (Hiratsuka, JP) |
Assignee: |
Lion Corporation (Tokyo,
JP)
|
Family
ID: |
15073522 |
Appl.
No.: |
06/748,393 |
Filed: |
June 24, 1985 |
Foreign Application Priority Data
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Jun 28, 1984 [JP] |
|
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59-132103 |
|
Current U.S.
Class: |
424/130.1;
424/49 |
Current CPC
Class: |
A61K
8/64 (20130101); A61Q 11/00 (20130101); A61K
9/0056 (20130101) |
Current International
Class: |
A61K
9/00 (20060101); A61K 039/395 () |
Field of
Search: |
;424/85,49,48,101
;260/112R,112B |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
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|
981260 |
|
Jan 1965 |
|
GB |
|
1492383 |
|
Nov 1977 |
|
GB |
|
1505513 |
|
Mar 1978 |
|
GB |
|
1545862 |
|
May 1979 |
|
GB |
|
2013691 |
|
Aug 1979 |
|
GB |
|
1573995 |
|
Mar 1980 |
|
GB |
|
Other References
Mouton et al., Infect, Immun., 31 (1), pp. 182-192 (1981)..
|
Primary Examiner: Hazel; Blondel
Attorney, Agent or Firm: Birch, Stewart, Kolasch &
Birch
Claims
What is claimed is:
1. A method for preventing Bacteroides gingivalis from colonizing
in the mouth comprising intraorally administering an effective
amount of a non-specific .gamma.-globulin to prevent Bacteroides
gingivalis from colonizing in the mouth.
2. The method as set forth in claim 1, wherein the nonspecific
.gamma.-globulin is one which is derived from a mammal.
3. The method as set forth in claim 1, wherein the nonspecific
.gamma.-globulin is human .gamma.-globulin.
4. The method as set forth in claim 1, wherein the method of
administering is selected from the group consisting of brushing,
gargling, massaging, and chewing.
5. The method as set forth in claim 1, wherein the non-specific
.gamma.-globulin is applied to the mouth of mammals selected from
the group consisting of humans, horses, cattle, goats, pigs, dogs
and cats.
6. The method as set forth in claim 1, wherein the dose of
.gamma.-globulin is 0.001 to 100 mg per kg per day.
7. The method as set forth in claim 1, wherein the dose of
.gamma.-globulin is 0.05 to 50 mg per kg per day.
8. The method as set forth in claim 1, wherein the composition is
applied to inflamed gingiva.
Description
BACKGROUND OF THE INVENTION
The present invention relates to a composition for oral application
to prevent and remedy the periodontal diseases such as gingivitis,
and more particularly, it relates to a composition for oral
application to prevent Bacteroides gingivalis, which is one of the
bacteria that cause periodontal diseases, from colonizing in the
mouth and thereby to prevent periodontal diseases.
There are many people having periodontal disease such as gingivitis
and periodontitis. The rate of such disease in adults is especially
on the increase. Prevention of periodontal diseases will be an
important problem under the future circumstance of an ever more
increasing number of aged persons.
Periodontal disease is primarily caused by bacteria existing in
accumulated plaque in periodontal pockets. A healthy periodontal
pocket is usually composed of an overwhelming amount of gram
positive bacteria, while the amount of gram negative bacteria
increases with the progress of the periodontal disease. Bacteroides
gingivalis, Fusobacterium nucleatum, Eikenella corrodens,
Actinobacillus actinomycetemcomitans and so forth are primarily
listed as such gram negative bacteria. In the focal regions of
adult patients with severe periodontal disease, gram negative
bacteria are detected in most cases among which Bacteroides
gingivalis is separated in a specially high frequency. In many of
these cases, the titer of anti-Bacteroides gingivalis antibody in
the serum of the patient also increases. In addition, it has been
demonstrated that the inoculation of an animal with Bacteroides
gingivalis aggravates periodontal inflammation. These results
indicate that Bacteroides gingivalis plays an important role in the
development of periodontal diseases.
Bacteroides gingivalis adheres to periodontal mucosa by means of
its pili and capsule existing on the surface of its bacterial body,
thereby proliferating and badly influencing the periodontal region.
For the prevention of periodontal diseases, the inhibition of the
colonization or suppression of the proliferation of Bacteroides
gingivalis in the mouth is effective and bactericides are mainly
used now to prevent periodontal disease. The use of a bactericide,
however, is not desirable because it indiscriminately kills
bacteria in the mouth and changes the bacterial flora in the mouth.
In some more specific methods, the inhibition of the colonization
of Bacteroides gingivalis in the mouth is attempted by using a
vaccine. However, since the whole bacterial cell is used as an
active vaccine directly injected into the living human body in all
of these methods, they have problems in terms of both effect and
toxicity.
SUMMARY OF THE INVENTION
An object of the present invention is to provide a composition for
oral application having an excellent effect on preventing
periodontal disease.
For the purpose of attaining the above object, the present
inventors conducted on a highly safe method of securing the
prevention of periodontal disease through effective suppression of
the colonization of the causative bacteria of periodontal disease,
especially Bacteroides gingivalis in the mouth. As a result, the
inventors have found that the oral administration of nonspecific
.gamma.-globulin is effective.
.gamma.-Globulin preparations have been known as being effective in
preventing and curing measles, asthma, varicella, and polio, and
they are used in the form of intramuscular injections. However, it
has not been known that .gamma.-globulin is effective against
periodontal diseases. Unexpectedly, the investigation by the
present inventors revealed that nonspecific .gamma.-globulin
prevents Bacteroides gingivalis from colonizing in the mouth when
it is intraorally administered, and that periodontal diseases can
be effectively prevented by blending nonspecific .gamma.-globulin
in a composition for oral application.
Therefore, the present invention provides a composition for oral
application comprising a nonspecific .gamma.-globulin as an
effective ingredient for suppressing the intraoral colonization of
Bacteroides gingivalis
According to this invention, since the composition contains the
nonspecific .gamma.-globulin, the colonization of Bacteroides
gingivalis in the mouth is effectively prevented, resulting in the
prevention of periodontal disease such as periodontitis.
In addition, since the nonspecific .gamma.-globulin is highly safe,
the composition according to this invention can be safely used.
The above and other objects, features, and advantages of this
invention will be more fully understood by reading the following
description.
DETAILED DESCRIPTION OF THE INVENTION
The composition for oral application of the present invention
contains a nonspecific .gamma.-globulin, and may be prepared and
used in various forms applicable to the mouth such as dentifrices
including toothpastes, toothpowders and liquid dentifrices, liquid
refrigerants including mouthwashes, solid refrigerants including
troches and chewing gums, dental pastes, gingival massage creams,
gargle tablets, dairy products including ice creams, yogurts and
babaroa bases, injections and the like. It may be applied to the
gingiva and other affected parts in the mouth by various methods
including brushing, gargling, massaging applying, chewing and
injecting. The scope of this invention covers those compositions
which are applied to the mouth of mammals such as horses, cattle,
goats, pigs, dogs, cats, and to the mouth of poultry.
According to this invention, the composition for oral application
is incorporated with nonspecific .gamma.- globulin as an effective
ingredient for the prevention and remedy of periodontal disease. In
the case where the composition is to be applied to a human, human
.gamma.-globulin may preferably be used, and the one for mammals
may preferably contain .gamma.-globulin derived from the same
species of animal to which it is applied. In addition, since
.gamma.-globulin is as safe as proteins in the feed,
.gamma.-globulin derived from the different species of animal to
which it is applied may also be used preferably. The nonspecific
.gamma.-globulin may be prepared by the well-known methods. For
example, it can be prepared from blood by the steps of separating
serum, performing salting-out with ammonium sulfate solution and
performing dialysis with water. It also can be prepared from milk
by the steps of centrifuging to collect the intermediate layer,
performing salting-out with ammonium sulfate solution and
performing dialysis with water. Commercially available
.gamma.-globulins can also be used.
The amount of .gamma.-globulin (dry basis) to be incorporated in
the composition varies depending on the kind of the composition. In
most cases, it is 0.001 to 10% by weight, particularly 0.01 to 1%
by weight of the composition, so that the dose of .gamma.-globulin
is 0.01 to 100 mg, preferably 0.05 to 50 mg, per kg per day. In the
case of injections, the preferred dose is 10 to 300 mg,
particularly 25 to 100 mg, per kg per day.
The composition according to this invention may further include
additional well-known ingredients depending on the type and form of
a particular composition. Any desired known ingredients may be
mixed with the nonspecific .gamma.-globulin.
In preparing dentifrice compositions, an abrasive may be blended
generally in an amount of 5 to 95%, especially 15 to 60% by weight
of the composition, including dicalcium phosphate dihydrate,
dicalcium phosphate anhydrate, monocalcium phosphate, tricalcium
tertiary phosphate, calcium carbonate, calcium pyrophosphate,
silica abrasives, aluminosilicate, aluminum oxide, aluminum
hydroxide, insoluble sodium metaphosphate, trimagnesium phosphate,
magnesium carbonate, calcium sulfate, titanium dioxide, resins, and
the like.
In preparing paste-like compositions, typically toothpastes, a
binder may be blended generally in an amount of 0.3 to 5% by
weight, including sodium carboxymethyl cellulose, methyl cellulose,
sodium carboxymethyl hydroxyethyl cellulose, hydroxyethyl
cellulose, sodium alginate, carrageenan, gum arabic, tragacanth
gum, karaya gum, polyvinylalcohol, sodium polyacrylate,
carboxyvinyl polymer, polyvinyl pyrrolidone, and the like.
In preparing paste-like and liquid oral compositions, typically
toothpastes and mouthwashes, a humectant may be blended generally
in an amount of 10 to 70% by weight, including polyethylene glycol,
ethylene glycol, sorbitol, glycerol, propylene glycol, 1,3-butylene
glycol, xylitol, maltitol, lactitol, and the like.
In addition to the above ingredients, a surface active agent
including water soluble salts of alkyl sulfate having 8 to 18
carbon atoms such as sodium laurate and sodium myristate, sodium
salts of higher fatty acids, water-soluble salts of sulfonates
monoglycerides of higher fatty acids having 10 to 18 carbon atoms
in the fatty acid group such as sodium lauryl monoglyceride
sulfonate and sodium coconut monoglyceride sulfonate, sodium
monoglyceride monosulfates of higher fatty acids, olefin
sulfonates, paraffin sulfonates, sodium N-methyl-N-palmitoyl
taurate, sodium N-lauroyl sarcosinate, sodium
N-lauroyl-.beta.-alaninate, stearyl monoglyceride, alkyrol ethanol
amides such as lauroyl monoethanol amide and lauroyl diethanol
amide, sucrose fatty acid esters having 12 to 18 carbon atoms in
the fatty acid group such as sucrose monolaurate and dilaurate,
lactose fatty acid esters, lactitol fatty acid esters, maltitol
fatty acid esters, stearic acid monoglyceride, polyoxyethylene
sorbitan monolaurate, polyoxyethylene-hardened castor oil,
condensates of sorbitan monostearate with approximately 60 moles of
ethylene glycol, condensates of ethylene oxide with propylene
oxide, and their derivatives such as polyoxyethylene
polyoxypropylene monolauryl ester, betaine and amino acid type
amphoteric surfactants, and the like may be blended in an amount of
0 to 10%, preferably 0.1 to 5%, more preferably 1 to 2.5% by weight
of the composition. A flavor such as an essential oil including
peppermint oil and spearmint oil and a flavoring material including
l-menthol, carvone, eugenol and anethole, a sweetener such as
sodium saccharinate, stevioside, neohesperidyldihydrochalcone,
glycyrrhizin, perillartine, p-methoxycinnamic aldehyde, a
preservative, and the like may be blended in an effective
amount.
In this invention, effective ingredients such as sorbic acid,
alexidine, hinokitiol, cetylpyridinium chloride, alkyl glycine,
alkyldiaminoethyl glycinate, allantoin, .epsilon.-aminocaproic
acid, tranexamic acid, azulene, vitamin E, a water soluble primary
or secondary phosphate, a quaternary ammonium compound, sodium
chloride, enzymes such as dextranase, mutanase, lytic enzyme and
protease, fluoride compounds such as sodium fluoride, stannous
fluoride and sodium monofluorophosphate, chlorhexidine and its
salts, bacteriocin, glucosyltransferase inhibitor, and crude drugs
and their extracts may also be blended in an effective amount.
Other types of compositions may also be prepared by selecting any
desired ingredients as usual and mixing them by a conventional
procedure.
Examples of the other ingredients for various types or forms of the
composition are shown in the following examples.
Paste-like and liquid oral compositions may generally have a pH
ranging from 5 to 10, but not limited thereto.
The composition for oral application of this invention may be
formulated into liquid that can be used for injections or powder
that can be prepared into injections upon application. The
composition in the form of injections may be incorporated with a
proper stabilizer (e.g., aminoacetic acid), preservative (e.g.,
thimerosal), and tonicity agent (e.g., sodium chloride) in the
usual way.
The composition of this invention may be applied in the known
manner according to the respective formulations. Preferably it
should be applied so that it acts on oral bacteria, and good
effects are resulted when it is applied to the inflamed gingiva for
periodontal disease. The composition in the form of injections may
be injected into the gingiva.
On account of nonspecific .gamma.-globulin incorporated therein as
mentioned above, the composition for oral application of this
invention prevents Bacteroides gingivalis from colonizing in the
mouth and thereby effectively prevents and remedies the
periodontosis such as gingivitis and periodentitis.
The invention is now described in more detail with reference to the
following examples, in which "%" means "% by weight" and the
quantity of .gamma.-globulin is expressed on dry basis.
EXAMPLE 1
(1) Preparation of .gamma.-globulin:
.gamma.-Globulin from blood
.gamma.-Globulin was prepared from blood of healthy rabbits by the
steps of separating serum, performing salting-out twice with 50%
ammonium sulfate solution, and performing dialysis with distilled
water according to a known method.
.gamma.-Globulin was prepared from goat milk by the steps of
centrifuging at 1500 rpm for 1 hour to collect the intermediate
layer, performing salting-out twice with 50% ammonium sulfate
solution, and performing dialysis with distilled water according to
a known method.
(2) Effect of rabbit .gamma.-globulin on preventing Bacteroides
gingivalis from colonizing in the hamster mouth.
Sixteen male golden hamsters arranged in test group and control
group, each group consisting of eight hamsters, were grown with
powder feed and free watering (tap water) until they were
slaughtered. On the first day, the hamsters in both groups were
treated as follows: The first molars on both sides of the lower jaw
were tied with a cotton thread (No. 50). Each molar was inoculated
with Bacteroides gingivalis 381-R' by dropping 0.1 ml of suspension
containing 1.times.10.sup.8 living bacteria per ml. Thirty minutes
later, the hamsters in test group were administered with a 50:50
mixture of nonspecific rabbit .gamma.-globulin and glycerin at a
dose of 0.05 ml for each molar, and then the rows of molars (both
the under side and the buccal side) were brushed 20 times in the
presence of the mixture. For the control group, on the other hand,
brushing was performed in the same way as mentioned above using a
50:50 mixture of distilled water and glycerin. The above procedure
was repeated during three consecutive days, and on the fourth day
and subsequent days, brushing alone was performed twice a day.
The cottom thread used for tying was recovered one week and three
weeks after the inoculation of living bateria, and the number of
Bateroides gingivalis and the total number of anaerobic bacteria in
the cotton thred were counted. Table 1 shows the number of hamsters
in which Bacteroides gingivalis colonized and the ratio of
colonization which is represented in percentage of the number of
Bacteroides gingivalis in the total number of anaeobic
bacteria.
TABLE 1 ______________________________________ Number of
colonization Ratio of colonization
______________________________________ After one week Test group
8/8 0.53 .+-. 0.20% Control group 8/8 1.07 .+-. 0.48% After three
weeks Test group 5/8 0.025 .+-. 0.041% Control group 7/8 0.951 .+-.
0.493% ______________________________________ Remarks: "Number of
colonization" is the number of hamsters in which Bacteroides
gingivalis colonized divided by the total number of hamsters
tested. "Ratio of colonization" is the number of Bacteroides
gingivalis divided b the total number of anaerobic bacteria
multiplied by 100 plus or minus SD
It is to be noted in Table 1 that colonization of Bacteroides
gingivalis was observed in five cases out of eight in the test
group three weeks after inoculation, and this is smaller than that
in the control group. In addition, the ratio of colonization was
apparently low in the test group when measured three weeks after
inculation. This indicates that the topical administration of
.gamma.-globulin prevents Bacteroides gingivalis from colonizing
and is effective against periodontal diseases.
EXAMPLE 2 (Toothpaste)
______________________________________ Dicalcium phosphate
dihydrate 50.0% Glycerin 20.0 Sodium carboxymethylcellulose 1.0
Sodium lauryl sulfate 1.5 Sodium lauroyl sarcosinate 0.5 Flavor 1.0
Sodium saccharin 0.1 Dextranase 0.01 Water Balance Total 100.0%
______________________________________
In addition to the above ingredients, the toothpaste was
incorporated with 0.01% or 0.05% of human .gamma.-globulin and
0.01% of chlorhexidine gluconate.
EXAMPLE 3 (Toothpaste)
______________________________________ Dicalcium phosphate
dihydrate 50.0% Sorbitol 10.0 Glycerin 10.0 Sodium
carboxymethylcellulose 1.0 Sodium lauryl sulfate 2.0 Flavor 1.0
Sodium saccharin 0.1 Ethanol 2.0 Mutanase 0.1 Water Balance Total
100.0% ______________________________________
In addition to the above ingredients, the toothpaste was
incorporated with 0.01% or 0.05% of human .gamma.-globulin, and
0.3% of sodium monofluorophosphate or 0.01% of chlorhexidine
gluconate or 0.05% of lytic enzyme or 0.02% of bacteriocin.
EXAMPLE 4 (Toothpaste)
______________________________________ Calcium carbonate 50.0%
Glycerin 20.0 Carrageenan 0.5 Sodium carboxymethylcellulose 1.0%
Lauroyl diethanolamide 1.0 Sucrose monolaurate 2.0 Flavor 1.0
Sodium saccharin 0.1 Water Balance Total 100.0%
______________________________________
In addition to the above ingredients, the toothpaste was
incorparated with 0.05% of human .gamma.-globulin, 0.005% of
chlorhexidine gluconate, and 0.05% of tranexamic acid.
EXAMPLE 5 (Toothpaste)
______________________________________ Dicalcium phosphate
dihydrate 50.0% Glycerin 20.0 Sodium carboxymethylcellulose 2.0
Sodium lauryl sulfate 2.0 Flavor 1.0 Sodium saccharin 0.1 Water
Balance Total 100.0% ______________________________________
In addition to the above ingredients, the toothpaste was
incorporated with 0.01% of horse .gamma.-globulin.
EXAMPLE 6 (Paste Toothpaste)
______________________________________ Silicic anhydride 30.0%
Glycerin 30.0 Sorbitol 20.0% Sodium carboxymethylcellulose 1.0
Sodium lauryl sulfate 2.0 Flavor 1.0 Sodium saccharin 0.1 Ethanol
2.0 Water Balance Total 100.0%
______________________________________
In addition to the above ingredients, the toothpaste was
incorporated with 0.02% of cattle .gamma.-globulin, and 0.1% of
sodium fluoride or 0.01% of chlorhexidine gluconate or 0.1% of
lytic enzyme or 0.01% of bacteriocin.
EXAMPLE 7 (Toothpaste)
______________________________________ Dicalcium phosphate
dihydrate 50.0% Calcium carbonate 30.0 Glycerin 10.0 .alpha.-Olefin
sulfonate 1.0 Flavor 1.0 Sodium saccharin 0.1 Dextran 0.5 Water
Balance Total 100.0% ______________________________________
In addition to the above ingredients, the toothpowder was
incorporated with 0.03% of human .gamma.-globulin, and 0.01% of
chlorhexidine gluconate or 0.05% of lytic enzyme or 0.001% of
bacteriocin.
EXAMPLE 8 (Liquid dentifrice)
______________________________________ Sodium polyacrylate 50.0%
Glycerin 30.0 Flavor 0.9 Sodium saccharin 3.0 Linolic acid 0.05
Water Balance Total 100.0%
______________________________________
In addition to the above ingredients, the liquid dentifrice was
incorporated with 0.01% of human .gamma.-globuloin, 0.05% of
chlorhexdine gluconate, and 0.05% of lytic enzyme or 0.001% of
bacteriocin.
EXAMPLE 9 (Mouthwash)
______________________________________ Ethanol 20.0% Flavor 1.0
Sodium saccharin 0.05 Lauroyl diethanolamide 0.3 Water Balance
Total 100.0% ______________________________________
In addition to the above ingredients, the mouthwash was
incorporated with 0.05% of human .gamma.-globulin and 0.01% of
chlorhexidine gluconate.
EXAMPLE 10 (Gargle tablet)
______________________________________ Sodium hydrogen carbonate
54.0 parts by weight Sodium secondary phosphate 10.0 parts by
weight Polyethylene glycol 3.0 parts by weight Citric acid 17.0
parts by weight Sodium sulfate (anhydrous) 13.6 parts by weight
Flavor 2.0 parts by weight Oleic acid 0.1 parts by weight
______________________________________
In additon to the above ingredients, the gargle tablet was
incorporated with 0.01% of human .gamma.-globulin and 0.05% of
chlorhexidine gluconate or 0.05% of lytic enzyme or 0.01% of
bacteriocin.
EXAMPLE 11 (Gingiva massage cream)
______________________________________ White vaselin 8.0% Propylene
glycol 4.0 Stearyl alcohol 8.0 Polyethylene glycol 4000 25.0
Polyethylene glycol 400 37.0 Sucrose stearate 0.5 Water Balance
Total 100.0% ______________________________________
In addition to the above ingredients, the cream was incorporated
with 0.05% of human .gamma.-globulin and 0.01% of chlorhexidine
gluconate or 0.1% of tocopherol acetate.
EXAMPLE 12 (Chewing gum)
______________________________________ Gum base 44.0% Calcium
carbonate 2.0 Corn syrup 15.0 Sugar 30.0 Sucrose palmitate 1.0
Fructose 4.0 Maltose 3.0 Flavor 1.0 Total 100.0%
______________________________________
In addition to the above ingredients, the chewing gum was
incorporated with 0.01% of human .gamma.-globulin and 0.01% of
chlorhexidine gluconate or 0.1% of lytic enzyme or 0.01% of
bacteriocin.
EXAMPLE 13 (Troche)
______________________________________ Gum arabic 6.0% Glucose 72.0
Gelatin 3.0 Flavor 0.2 l-Menthol 0.1 Spearmint oil 0.1 Sodium
ascorbate 0.1 Water Balance Total 100.0%
______________________________________
In addition to the above ingredinets, the troche was incorporated
with 0.001% of human .gamma.-globulin and 0.01% of chlorhexidine
gluconate or 0.05% of lytic enzyme or 0.01% of bacteriocin or 0.05%
of tocopherol nicotinate.
EXAMPLE 14 (Buccal paste)
______________________________________ Polyoxyethylene monostearate
2.0% Sorbitan monooleate 2.0 Myristyl alcohol 2.0 Palmityl alcohol
3.0 Propylene glycol 15.0 Sodium carboxymethylcellulose 5.0 Gelatin
1.0 Sodium saccharin 0.2 Peppermint oil 0.5 Spearmint oil 0.5
Lysozyme chloride 5000 units/g Water Balance Total 100.0%
______________________________________
In additon to the above ingredients, the paste was incorporated
with 0.001% or 0.05% of human .gamma.-globulin and 0.1% of stannous
fluoride or 0.01% of chlorhexidine hydrochloride or 0.05% of lytic
enzyme or 0.01% of bacteriocin.
EXAMPLE 15 (Buccal paste)
______________________________________ Glyceryl monostearate 3.0%
Oleyl alcohol 5.0 Polyethylene glycol 15.0 White vaselin 3.0
Monosodium N--palmitoylglutamate 0.5 Hydroxyethylcellulose 5.0
Tocopherol acetate 0.1 Sodium saccharin 0.2 Mentha oil 0.7 Carvone
0.5 Anethole 0.3 Eugenol 0.1 Water Balance Total 100.0%
______________________________________
In addition to the above ingredients, the paste was incorporated
with 0.05% of human .gamma.-globulin, 0.01% of chlorhexidine
hydrochloride, and 0.1% of .beta.-glycyrrhetinic acid.
EXAMPLE 16 (Ice cream)
______________________________________ Cream (fat 50%) 16.84% Cow's
milk (fat 3.7%) 42.67 Evaporated condensed skim milk 24.24 Sugar
11.25 Corn syrup 4.65% Stabilizer 0.35 Total 100.0%
______________________________________
In addition to the above ingredients, the ice cream was
incorporated with 0.1% of cattle .gamma.-globulin.
EXAMPLE 17 (Ice cream)
______________________________________ Cream (fat 40%) 31.54% Cow's
milk (fat 3.7%) 37.16 Evaporated condensed skim milk 15.08 Sugar
11.25 Corn syrup 4.67 Stabilizer 0.30 Total 100.0%
______________________________________
In addition to the above ingredients, the ice cream was
incorporated with 0.5% of cattle .gamma.-globulin.
EXAMPLE 18 (Yoghurt)
______________________________________ Skim milk 320 kg Sweetened
condensed milk 56 Sugar 23
______________________________________
Yoghurt was produced by fermentation in the usual way.
EXAMPLE 19 (Bavaroise dough)
______________________________________ Cow's milk 370 cc Sugar 60 g
Vanilla sheath 1 piece Egg yolk 5 pieces Gelatin 20 g Raw cream 75
cc ______________________________________
In addition to the above ingredients of Examples 18 and 19, 0.5% of
cattle .gamma.-globulin were incorporated.
EXAMPLE 20
Wister male rats arranged in test group and control group, each
consisting of 10 rats, were grown with Keyes diet 2000 for 12 weeks
(112 age in day) from weaning (at 28 age in day) to slaughter. The
rats in test group were given twice each of .gamma.-globulin in
(made by Miles Laboratories, Inc. and Cappel Laboratories, Inc.) by
injection into the tail vein at 84 age in day (12 age in week) and
109 age in day (3 days before slaughter). The rats in control group
were given twice 1 ml each of sterillized phosphate buffer solution
on the same age in day as above.
Immediately after injection of .gamma.-globulin in or phosphate
buffer solution and immediately before slaughter, blood was
collected from the tail vein or orbit. After standing at 4.degree.
C. for 30 minutes, the blood was centrifuged at 2500 rpm for 10
minutes to separate serum. The serum was stored at -70.degree.
C.
Immediately after slaughter with ether, the upper and lower jaws
were washed with physiological saline solution and fixed with 10%
formalin. After deliming with Plank-Rychlo deliming solution, block
sections were prepared for mesiodistal sectional specimens.
The specimen was embedded in paraffin according to Heeley method
and 8-.mu.m thick sections were prepared, followed by dyeing with
hematoxylin-eosin double stain. For each rat, four sections of the
upper jaw and four sections of lower jaw were examined for the
number of polymorphonuclear and mononuclear infiltration cells at
each mamillary body. By using eight specimens for each rat, the
ratio of the infiltration region to the mamillary body connective
tissue in the area from the cement-enamel boundary to the tooth
crown was measured with a planimeter according to Line method. The
results are shown in Table 2.
TABLE 2 ______________________________________ Number of
Infiltration Rat No. infiltration cells area (%)
______________________________________ Test group 1 177 24.0 2 321
38.5 3 65 22.5 4 219 18.5 5 148 26.8 6 228 34.3 7 500 38.0 8 181
14.3 9 273 17.0 10 374 20.8 Average 249 .+-. 39 25.5 .+-. 2.8
Control group 11 175 19.8 12 298 19.0 13 154 31.3 14 159 25.0 15
159 58.0 16 345 35.3 17 451 24.0 18 344 34.2 19 446 31.3 20 271
38.3 Average 280 .+-. 37 31.6 .+-. 3.6
______________________________________
It is to be noted from Table 2 that the number of infiltration
cells and the infiltration area are lower in test group (which was
given .gamma.-globulin) than in control group. This result suggests
that .gamma.-globulin is effective against periodontal
diseases.
It is known that the cellular infiltration of leukocytes and the
decomposition of connective tissue components take place in the
early stage of periodontal diseases, and the disintegration of
connective tissues and the extreme infiltration of plasma cells
take place in the matured stage. This suggests that suppressing the
infiltration of cell is effective against periodontal diseases. It
was observed in this example that the infiltration of gingival
cells is effectively suppressed by injection of
.gamma.-globulin.
EXAMPLE 21
An injection of the following formula was prepared.
______________________________________ Human .gamma.-globulin 15%
Aminoacetic acid 2.25% Methylol 0.01% Distilled water for injection
Balance Total 100.0% pH 6.4 to 7.2
______________________________________
For the prevention and remedy of human marginal periodontosis of
patient at P.sub.1 and P.sub.2 levels, this injection is injected
into the inflamed root of a tooth at a dose of 50 mg of
.gamma.-globulin (equivalent to 0.33 ml of the injection) per kg of
body weight after scaling in order to alleviate inflammation and to
prevent the resorption of the alvolar bone. For patients at P.sub.3
level, it is injected in the same manner after scaling at a dose of
75 mg of .gamma.-globulin (equivalent to 0.50 ml of the injection)
per kg of body weight.
For the prevention and remedy of dog's and cat's periodontosis, an
injection of the same formula as above (except the type of
.gamma.-globulin is different) is injected into the inflamed root
of a tooth at a dose of 25 mg of .gamma.-globulin per kg of body
weight. In either cases, good results were obtained.
* * * * *